Herzig, Maryanne C. S. published the artcileSynthesis and characterization of N-hydroxysuccinimide ester chemical affinity derivatives of asialoorosomucoid that covalently crosslink to galactosyl receptors on isolated rat hepatocytes, Related Products of alcohols-buliding-blocks, the publication is Biochemistry (1989), 28(2), 600-10, database is CAplus and MEDLINE.
Asialoorosomucoid (ASOR) was derivatized with 5 homobifunctional N-hydroxysuccinimide (NHS) ester crosslinkers. NHS/ASOR derivatives were synthesized, purified, and applied within 10 min to isolated rat hepatocytes at 4°. Specific binding of these 125I-labeled derivatives was ∼90% in the presence of either EGTA or excess ASOR. Specific crosslinking, assessed by the resistance of specifically bound NHS/125I-ASOR to release by EGTA, was 50-75% of the specifically bound ligand. The extent of specific crosslinking correlated with the average number of NHS groups per ASOR and was controlled by varying the molar ratio of crosslinker to ASOR during the synthesis. Crosslinking proceeded rapidly at 4° as a 1st-order process (k = 0.25 min-1, t1/2 = 2.8 min). After being crosslinked with any of the NHS/125I-ASOR derivatives, cells were washed with EGTA, solubilized in Triton X 100, and analyzed by SDS-PAGE and autoradiog. Major bands were observed at Mr ≃ 83, 94, and 105 kilodaltons corresponding to the expected size of 1:1 adducts between NHS/ASOR (Mr ≃ 41.3 kilodaltons) and the 3 subunits of the receptor, Mr ≃ 43, 50, and 60 kilodaltons. The 3 subunits, rat hepatic lectin (RHL) 1, 2, and 3, were labeled in the ratio of about 1.0:1.2:1.0, resp. After crosslinking, a polyclonal goat antibody to the receptor immunoprecipitated up to 100% of the specifically crosslinked NHS/125I-ASOR. Preimmune IgG immunoprecipitated <1% of the radiolabeled ligand. Cell surface receptors were crosslinked to NHS-ASOR, extracted with Triton X 100, immunoprecipitated with anti-orosomucoid-Sepharose, and subjected to Western blot anal. By use of antisera specific for RHL 1 or RHL 2/3 (from K. Drickamer), crosslinked complexes of Mr ∼85 kilodaltons or ∼90-115 kilodaltons, resp., were detected as were uncrosslinked native subunits. The ratio of free to crosslinked subunits was ∼10:1 for RHL 1 and ∼0.5:1 for RHL 2/3. Apparently, all 3 receptor subunits can crosslink to ligand. A model is proposed in which the native receptor is a heterohexamer composed of 4 subunits of RHL 1 and 2 subunits of RHL 2 and/or RHL 3.
Biochemistry published new progress about 70539-42-3. 70539-42-3 belongs to alcohols-buliding-blocks, auxiliary class pyrrolidine,Ester,Amide,Inhibitor,Inhibitor, name is Bis(2,5-dioxopyrrolidin-1-yl) O,O’-ethane-1,2-diyl disuccinate, and the molecular formula is C18H20N2O12, Related Products of alcohols-buliding-blocks.
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